Hepatogenic Differentiation of Human Induced Pluripotent Stem cells on Collagen-Coated Polyethersulfone Nanofibers

Abstract

Many scientists have been fascinated with induced pluripotent stem cells (iPSCs) for cell replacement therapies. Nanofibrous biocompatible scaffolds have been shown to foster better cell adhesion and improve stem cell differentiation. In the current study, after fabrication using electrospinning technique and surface modifications, the characteristics of polyethersulfone (PES) nanofibers were determined by scanning electron microscopy, attenuated total reflection Fourier transform infrared spectroscopy, and 3-[4, 5-dimethylthiazol-2-yl]-2, 5 diphenyltetrazolium bromide (MTT) assay. Then, the hepatogenic potential of iPSCs was evaluated using real-time reverse transcription polymerase chain reaction (RT-PCR) and immunocytochemistry (ICC) after culture on collagen-coated polyethersulfone (PES/COL) scaffolds. After scaffolds characterization, analysis of two important definitive endoderm specific markers (Sox17 and Foxa2) using real-time RT-PCR and ICC indicated increase in their mRNA and protein levels after 5 days of hepatogenic induction. In addition, to determine hepatic differentiation of iPSCs cultured on PES/COL, the expression of albumin and α-fetoprotein was evaluated by ICC after 20 days. Real-time RT-PCR analysis showed increased expression of albumin, TAT, cytokeratin 19, and Cyp7A1 genes during the course of differentiation program. Finally, enzyme-linked immunosorbent assay analysis demonstrated an increased expression of albumin in the protein level after 28 days of differentiation. In conclusion, our results demonstrated that PES/COL nanofibrous scaffolds could be a proper substrate to significantly increase the hepatogenic differentiation potential of iPSCs and could also be introduced as a promising candidate for liver tissue engineering applications.

Three-dimensional nanofiberous PLLA/PCL scaffold improved biochemical and molecular markers hiPS cell-derived insulin-producing islet-like cells

“…As well as in line with enderami and mansuor in 3 D culture, these markers were significantly more expressed than cells which were induced in two dimentional. Similar to our finding, Mahmoodinia et al [38] performed differentiation of human iPSCs on IPCs on hybrid nanofibrous scaffold and they showed successful expression of protein marker C-peptide in IPS Cell-derived Insulin-Producing Islet-Like cells.…”

Hybrid poly‐l‐lactic acid/poly(ε‐caprolactone) nanofibrous scaffold can improve biochemical and molecular markers of human induced pluripotent stem cell‐derived hepatocyte‐like cells

“…But the expression level of ALB and CK‐18 in cells cultured on PLLA scaffold in her study was significantly lower than our result, in which cells differentiated on PLLA/PCL membrane, this inconsistency may be due to the type of stem cell and the nanofiber used. Additionally, Mahmoodinia Maymand et al () have demonstrated that the expression level of marker ALB in hepatocyte differentiated on PES is lower than our result that is five‐folds. These results show that hiPSCs may be better differentiated on hybrid scaffold than single PES nanofibrous membrane.…”

“…Nanofibrous scaffolds are synthetic biocompatible scaffolds which have topographic structures that can be fabricated to mimic the structures of natural ECM (Mashhadikhan, Soleimani, Parivar, & Yaghmaei, ). Preliminary studies have used various scaffolds such as, polyethersulfone (PES; Hashemi et al, ; Kazemnejad et al, ; Mahmoodinia Maymand et al, ), poly‐ l ‐lactic acid (PLLA; Feng et al, ; Ghaedi, Soleimani, Shabani, Duan, & Lotfi, ), and PCL/collagen/PES (Mahmoodinia Maymand et al, ) for hepatocyte differentiation. However, these scaffolds have limitations, for example, Y.…”

Collagen coated electrospun polyethersulfon nanofibers improved insulin producing cells differentiation potential of human induced pluripotent stem cells

“…Electrospun PES nanofibrous scaffold was according to a protocol modified with Shabani et al [26,27] and after plasma treatment [28], more surface modification was performed by a coating of the collagen on the surface of the fabricated PES nanofibers. Collagen solution (0.1 ng/ml, Sigma Aldrich, St. Louis, MO) was added to the nanofibers and incubated at 4 C overnight and later washed two times by PBS.…”